Persimmon Research Articles

Influence of the extraction conditions on the carbohydrate and phenolic composition of functional pectin from persimmon waste streams

Persimmon fruit (Diospyros kaki Thunb.) production suffers great losses (15–20%) due to the inefficient overripening process. In order to valorise this waste, a compositional characterization of different fruit stages of the residues was done, and immature fruit was selected due to its high pectin and very high polyphenol content. A 3-level full factorial design was carried out to study the effect of temperature (70–95 °C) and low pH (0.5–1.5), on yield, degree of esterification, carbohydrate constituents, phenolic content and antioxidant capacity of the extracted pectin, and a complete polyphenol profile (UPLC-MS/MS) was performed on selected extracts. All responses could be accurately adjusted to the models (R2 > 80; lack of fit). Pectin yield, phenolic compounds and antioxidant activity ranged from 1.4 to 4.5%, 53.3 ± 2.27 to 111.7 ± 9.74 mg GAE/g pectin and 0.29 ± 0.01 to 2.77 ± 0.04 TEAC (Trolox μmol/mg pectin), respectively. A strong pectin-polyphenol interaction was found, which significantly enhanced acid resistance of both the pectin and polyphenol constituents, with optimum yield and polyphenol content at pH 1 and 95 °C. These new pectin-based ingredients might have a great potential as functional foods or natural food ingredients enhancing the quality and shelf-life.

Low intensity of high pressure processing increases extractable recovery of polyphenols and antioxidant activities of non-astringent persimmon fruit

This study aimed to investigate the effect of low intensity of high pressure processing (LHP) on bioactive compound content and the antioxidant capacity of persimmon before digestion and during simulated in vitro gastrointestinal digestion (IVD). The astringent and non-astringent persimmon cultivars treated with LHP at 100 MPa for 10 min were analyzed for polyphenols, antioxidant activities and bioaccessibility. The results showed that LHP treatment significantly increased (P < 0.05) the extractability of polyphenols and antioxidant activities of non-astringent persimmon by 1.5–2.0 folds as compared to the non-treated control treatment. Additionally, LHP exhibited positive results for non-astringent cultivar as indicated by the higher recovery of polyphenols (204–279%), bioaccessibility index (192–205%) and residual antioxidant activities (44–399%) in comparison to the control after IVD. However, LHP treatment did not affect astringent cultivar. These findings suggest that the LHP could be applied to enhance nutritional property of non-astringent persimmon and may be used for developing novel health products with high functional bioactive compounds which tolerate the digestion system.

Drying of persimmon fruit (Diospyros kaki L.) pretreated by different osmotic processes

AbstractPersimmon is a short shelf life fruit with interesting potential for application in different industrial segments. The immersion of food in a hypertonic (30 and 45 °Brix) or even hypotonic medium can be used as a pretreatment for the drying process. In this study, the convective drying of persimmons was preceded by osmotic dehydration (OD) with fructose in a standard atmosphere, pulsed vacuum osmotic dehydration (PVOD), ultrasound (US)‐assisted OD, and US pretreatment in distilled water. Pretreatment times were 0, 10, and 30 min. The samples were subjected to convective drying at 40 and 60°C, 0.5 m s−1. The effects of the pretreatments on the drying kinetics, volumetric shrinkage, and rehydration capacity were investigated. Fick's model and the Dincer and Dost model were used to fit the drying evolution. In addition to the influence of the osmotic solution concentration, moisture content and solid gain were functions of the treatment type. Convective drying required 260–740 min. The use of PVOD and US pretreatments resulted in the shortest drying times. The dried fruits showed particular characteristics with each different pretreatment. Both tested models presented satisfactory adjustments, but Fick's model was the best approach. In general, the treatments caused structural modifications in the persimmon fruits that modified the drying kinetics and the properties of the dried food.Practical ApplicationsConvective drying is a food processing method used to extend the shelf life of fruits, such as persimmons. The use of pretreatments is an interesting strategy to improve the drying process. However, the combination of these technologies can influence drying kinetics. Therefore, the use of mathematical models is important to represent the behavior of these processes.

Niastella soli sp. nov., isolated from rhizospheric soil of a persimmon tree.

A Gram-stain-negative, aerobic, motile and rod-shaped novel bacterial strain, designated MAH-29T, was isolated from rhizospheric soil of a persimmon tree. The colonies were light pink coloured, smooth, spherical and 0.1-0.8 mm in diameter when grown on Reasoner's 2A (R2A) agar for 2 days. Strain MAH-29T was able to grow at 20-37 °C, at pH 5.0-8.5 and at 0-2.0 % NaCl. Cell growth occurred on nutrient agar and R2A agar. The strain was positive in both oxidase and catalase tests. According to the 16S rRNA gene sequence comparisons, the isolate was identified as a member of the genus Niastella and was closely related to Niastella vici DJ57T (97.7 % similarity), Niastella koreensis GR20-10T (97.1 %) and Niastella yeongjuensis GR20-13T (97.0 %). Strain MAH-29T has a draft genome size of 8 876 333 bp (31 contigs), annotated with 6920 protein-coding genes, 61 tRNA and four rRNA genes. The average nucleotide identity and digital DNA-DNA hybridization values between strain MAH-29T and three closely related type strains were in the range of 78.2-83.2 % and 22.1-27.0 %, respectively. The genomic DNA G+C content was 43.8 mol%. The predominant isoprenoid quinone was menaquinone 7. The major fatty acids were identified as iso-C15:0, iso-C15:1 G and iso-C17:0 3OH. On the basis of DNA-DNA hybridization results, genotypic analysis and chemotaxonomic and physiological data, strain MAH-29T represents a novel species within the genus Niastella, for which the name Niastella soli sp. nov. is proposed, with MAH-29T (=KACC 19969T=CGMCC 1.16606T) as the type strain.

Extraction optimization and structural characterization of pectin from persimmon fruit (Diospyros kaki Thunb. var. Rojo brillante)

In this work we have efficiently extracted and characterized pectin from different tissues of astringent (AS) and non-astringent (NAS) persimmon fruits (peel, pulp, whole fruit) for the first time. The highest pectin extraction (≥7.2%) was carried out at 80 °C, 120 min with 1.5% sodium citrate in peel of both AS and NAS persimmon samples. All persimmon pectins showed a molecular weight and galacturonic acid content upper than 328 kDa and 78%, respectively, indicating their suitability as food ingredient. Pectin extracted from AS pulp and peel tissues exhibited an enriched structure in rhamnose and arabinose, whereas the opposite behavior was observed in NAS persimmon whole fruit samples. Remarkably, both pulp tissues (AS and NAS) presented the highest levels of glucose and mannose, non-pectic carbohydrates. In addition, techno-functional assessment (zeta potential, particle size, apparent viscosity, gelation) showed the suitability of the persimmon pectins for a broad range of industrial applications.

Chitosan-Phenylalanine Nanoparticles (Cs-Phe Nps) Extend the Postharvest Life of Persimmon (Diospyros kaki) Fruits under Chilling Stress

There are high levels of damage imposed on persimmon fruit postharvest, especially after storing it in cold storage, which causes chilling injury (CI). To reduce this stress on the fruit, the conventional way is to use chemical treatments. Since there is a limitation in the use of chemical materials, it is necessary to apply non-harmful treatments to decrease chilling injury and maintain the quality of persimmon in cold storage. The aim of this study is to investigate the effects of chitosan-loaded phenylalanine nanoparticles (Cs-Phe NPs) (2.5 and 5 mM) on physiochemical and quality factors of persimmon (Diospyros kaki) during 45 days of storage at 4 °C (38 °F) and evaluate the impact of Cs-Phe NPs on the preserving quality in order to reduce the chilling injury of this fruit. The experiment was conducted using a completely randomized design with three replications. Treatments were applied at 15, 30, and 45 days after storage at 4 °C with ≥90% relative humidity. The size of Cs-Phe NPs was less than 100 nm, approximately. The results showed that application of 5 mM of Cs-Phe NPs delayed the negative effects of chilling stress and enhanced antioxidant capacity, firmness, and total soluble solids of persimmon fruit. Lower H2O2 and malonaldehyde (MDA) accumulation along with higher soluble tannin and total carotenoid accumulation in persimmon fruit treated with 5 mM Cs-Phe NPs was also observed. Fruit coated using Cs-Phe NPs in both concentrations (2.5 and 5 mM) showed the highest antioxidant enzyme activity for superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX) and the lowest for polyphenol oxidase (PPO) and chilling injury during storage. According to our results, 5 mM of Cs-Phe NPs could be considered as the best treatment under chilling-stress conditions.

English

Mealybugs are agricultural pests occurring all over the world. The Sao Francisco Valley is one of the most productive regions regarding the cultivation of fruit crops in Brazil, demanding a proper phytosanitary management that can be obtained through the biological control of pests. Fortnight samples of mealybugs associated with roots, trunk, leaves and fruits of grapevines, pear, apple, persimmon, guava and Barbados cherry were performed in commercial farms in the sub-medium Sao Franciso Valley, Brazil, during twelve months. In order to identify mealybugs based on morphological characteristics from adult female specimens, samples were stored in Eppendorf tubes containing ethanol 70%, labeled and then identified. A total of 10,189 mealybugs, including nymphs and adult females were collected, with the highest number of specimens found in grapevines. The second most affected crop was persimmon, followed by pear. The lowest infestation was observed in guava, apple and Barbados cherry. Pseudococcidae evidenced preference for fruits. The following mealybugs were observed in the sub-medium region of the Sao Francisco Valley: Planococcus sp., Planococcus citri, Phenacoccus solenopsis, Dysmicoccus brevipes, Maconellicoccus hirsutus, Ferrisia virgata and Pseudococcus elisae. Key words: Fluctuation, identification, morphology, pests.

Traits and Gene Expression Analysis of Early and Late Ripening Pollination Constant Non-Astringent Persimmon Fruits at Different Developmental Phases

Traits and Gene Expression Analysis of Early and Late Ripening Pollination Constant Non-Astringent Persimmon Fruits at Different Developmental Phases

Transcription factors DkBZR1/2 regulate cell wall degradation genes and ethylene biosynthesis genes during persimmon fruit ripening.

BRASSINAZOLE RESISTANT (BZR) transcription factors are critical components of the brassinosteroid signalling pathway, but their possible roles in fruit ripening have rarely been reported. In this study, four BZR sequences were isolated from persimmon fruit. Among the four BZR genes, DkBZR1/2 were expressed in persimmon fruit; DkBZR1 protein amount decreased and dephosphorylated DkBZR2 gradually accumulated during the storage period. DkBZR1/2 proteins were localized in both the nucleus and cytoplasm and accumulated in the nucleus after 24-epibrassinolide treatment. DkBZR1 suppressed the transcription of Diospyros kaki endo-1,4-betaglucanase 1 (DkEGase1) and 1-aminocyclopropane-1-carboxylate synthase 1 (DkACS1) by binding to the BR response element (BRRE) in their promoters, and DkBZR2 activated the transcription of pectate lyase 1 (DkPL1) and 1-aminocyclopropane-1-carboxylate oxidase 2 (DkACO2) by binding to the E-box motif in their promoters. Transient overexpression of DkBZR2 promoted the conversion of acid-soluble pectin to water-soluble pectin and increased ethylene production in persimmon fruit. Our findings indicate that DkBZR1 and DkBZR2 serve as repressors and activators of persimmon fruit ripening, respectively.

Effects of Drip Irrigation Design on a Lemon and a Young Persimmon Orchard in Semi-Arid Conditions

Drip irrigation is presently widely recognized as the most efficient irrigation system that can be used in woody perennial crops. However, uncertainties exist on the more appropriate agronomic design to employ. Here, we summarized the research carried out for three seasons in two young woody perennial crops (persimmon and lemon) in southeastern Spain. Several irrigation designs were compared by maintaining a similar amount of water application but varying the number of emitters and pipelines in each row in the orchard. In the lemon trial, the agronomic irrigation design was additionally combined with different irrigation regimes, comparing full irrigation (FI) with sustained deficit irrigation (SDI). In the persimmon trees, which were still at the juvenility stage, varying the number of emitters per tree or the number of drip lines per tree row, neither affects tree performance nor fruit yield in two out of the three seasons. However, over the entire experimental period, the relative trunk growth increased when more emitters were employed. In the lemon trial, carried out with trees that had reached commercial production, the FI, compared with SDI, increased trunk growth and average fruit weight, while a reduced number of fruits per tree without affecting total yield was observed in the third year of experimentation. The number of emitters per tree only had an effect the first year, increasing lemon fruit weight when the number of drippers per tree increased. In addition, fruit composition was not consistently affected by the irrigation design. It is concluded that, for a given irrigation dose, irrigation frequency, and soil conditions (loam-clay texture), in both very young and more mature trees, increasing the number of emitters or the wetted area only had some slight positive effects on tree performance.

Analysis of morphological characteristics and phenetic relationship of ebony (Diospyros spp.) in Indonesia

Abstract. Rindyaastuti R, Hapsati L, Wibowo AT. 2021. Analysis of morphological characteristics and phenetic relationship of ebony (Diospyros spp.) in Indonesia. Biodiversitas 22: 2738-2753. Diospyros L. (Ebenaceae) is an economically important genus that covers over 500 plant species. Members of this group are known to produce hard, dark, high-quality timbers known as ebony trees, while other members are known as persimmon trees. There is limited information on the morphological characters and phenetic relationship of this genus from Indonesian archipelago. In this work, we reported the phenetic clustering of 14 species of Indonesian Diospyros, based on plant habit, stem, leaves, and fruit characters, which was analysed using UPGMA and Jaccard similarity index. The phenetic dendrogram divided the 14 species of Diospyros into two main clusters which separating Subgen. Maba and Eudiospyros. Further, subgen. Maba was clustered into a single cluster while subgen. Eudiospyros was divided into 3 sub-clusters as sister groups. The clustering in Eudiospyros was supported by low SI and bootstrap value, demonstrating the high morphological variation of the subgenus. The sectional separation of Diospyros spp. was considered as paraphyletic. In general, our phenetic clustering exhibited suitability and relevancy with previous classification. Fruit size and trichomes are very important in this phenetic clustering, especially to form Eudiospyros subgenus. Fruit trichomes were also special characters related to biogeographical distribution following the Wallace line theory.

Phenotypic Characterization of Postharvest Fruit Qualities in Astringent and Non-astringent Persimmon (Diospyros kaki) Cultivars.

Phenotypic characterization of postharvest traits is essential for the breeding of high-quality fruits. To compare postharvest traits of different genetic lines, it is essential to use a reference point during fruit development that will be common to all the lines. In this study, we employed a non-destructive parameter of chlorophyll levels to establish a similar physiological age and compared several postharvest traits of ten astringent and seven non-astringent persimmon cultivars. The fruit’s traits examined were astringency, weight, total soluble solids (TSS), titratable acidity (TA), chlorophyll levels (IAD), color (hue), firmness, color development and firmness loss during storage, crack development, and susceptibility to Alternaria infection. Although the chlorophyll (IAD) index and color (hue) showed a high correlation among mature fruits of all cultivars, the chlorophyll parameter could detect higher variability in each cultivar, suggesting that IAD is a more rigorous parameter for detecting the developmental stage. The average weight, TSS, and TA were similar between astringent and non-astringent cultivars. Cracks appeared only on a few cultivars at harvest. Resistance to Alternaria infection and firmness were lower in astringent than in non-astringent cultivars. Only the astringent cultivar “32” was resistant to infection possibly due to the existence of an efficient peel barrier. It was concluded that a high correlation existed between astringency, susceptibility to Alternaria infection, and firmness. Cracks did not correlate with astringency or firmness. The phenotypic traits evaluated in this work can be used in future breeding programs for elite persimmon fruits.

Tannin extraction and changes in content during the maturation of Mopan persimmon fruits

Tannin extraction and changes in content during the maturation of Mopan persimmon fruits

Lycopene β-cyclase plays a critical role in carotenoid biosynthesis during persimmon fruit development and postharvest ripening

The aim of the current study was to characterize the functional role of DkLCYb in carotenoid biosynthesis in persimmon fruit. The accumulation of carotenoids and the expression of carotenogenic genes were investigated during persimmon fruit development and postharvest ripening. DkLCYb was cloned and its functional role in carotenoid biosynthesis was characterized. The results showed that carotenoids constantly accumulated during persimmon fruit development and postharvest ripening. More carotenoids accumulated during postharvest storage than during fruit development. The carotenoid composition of unsaponified extracts of the persimmon fruit flesh showed that the major carotenoids were lycopene and β-carotene in ‘Huoguan’ and ‘Heishi’, respectively. Consistent with this result, the expression of six carotenogenic genes increased to their highest levels during postharvest storage. The selected carotenoid biosynthesis genes showed an increased expression pattern during fruit development and in the early period of postharvest storage. A significant decrease in carotenogenic genes was found after stage 6 (DkPSY, DkLCYb, DkZDS and DkBCH) or stage 5 (DkLCYe,). Both DkLCYe and DkBCH showed a steady increase from stage 1 to stage 5; however, dramatically decreased expression of DkLCYe and sharply increased expression of DkBCH were found at stage 6. Correlation analysis showed that the expression of DkPSY, DkPDS, DkZDS, DkLCYb and DkBCH was significantly correlated with the accumulation of one or more carotenoid compounds analysed in this study. However, a negative correlation was found between the expression of DkLCYe and the accumulation of lycopene, β-cryptoxanthin and zeaxanthin. Specifically, expression of DkLCYb showed the highest correlation (p < 0.01) with β-carotene (r = 0.74) accumulation. We cloned DkLCYb and analysed its functions in vitro and in vivo. DkLCYb contained typical domains that were highly conserved and essential for lcyb protein activity. The cyclization activity of the encoded protein was demonstrated by functional analysis in E. coli BL21. Transient overexpression of DkLCYb significantly enhanced the accumulation of β-carotene in persimmon fruit. These results indicate that DkLCYb is a genuine lycopene cyclase gene and it plays an essential role in β, β-carotenoid (β-carotene and β-cryptoxanthin) biosynthesis in persimmon fruit.

DkMYB14 is a bifunctional transcription factor that regulates the accumulation of proanthocyanidin in persimmon fruit

SUMMARYProanthocyanidins (PAs) are phenolic secondary metabolites that contribute to the protection of plant and human health. Persimmon (Diospyros kaki Thunb.) can accumulate abundant PAs in fruit, which cause a strong sensation of astringency. Proanthocyanidins can be classified into soluble and insoluble PAs; the former cause astringency but the latter do not. Soluble PAs can be converted into insoluble PAs upon interacting with acetaldehydes. We demonstrate here that DkMYB14, which regulates the accumulation of PA in persimmon fruit flesh, is a bifunctional transcription factor that acts as a repressor in PA biosynthesis but becomes an activator when involved in acetaldehyde biosynthesis. Interestingly, both functions contribute to the elimination of astringency by decreasing PA biosynthesis and promoting its insolubilization. We show that the amino acid Gly39 in the R2 domain and the ethylene response factor‐associated amphiphilic repression‐like motif in the C‐terminal of DkMYB14 are essential for the regulation of both PA and acetaldehyde synthesis. The repressive function of DkMYB14 was lost after the mutation of either motif, and all activities of DkMYB14 were eliminated following the mutation of both motifs. Our results demonstrate that DkMYB14 functions as both a transcriptional activator and a repressor, directly repressing biosynthesis of PA and promoting its insolubilization, resulting in non‐astringency in persimmon.

Effects of 1-methylcyclopropene and aminoethoxyvinylglycine treatments on fruit quality and antioxidant metabolites in cold-stored ‘Sangjudungsi’ persimmons

After harvest, ‘Sangjudungsi’ persimmon (Diospyros kaki Thunb.) fruit quickly soften and develop physiological disorders during cold storage. The objective of this study was to evaluate the effectiveness of 1-methylcyclopropene (1-MCP) and aminoethoxyvinylglycine (AVG) treatments on fruit quality attributes and antioxidant metabolites in ‘Sangjudungsi’ persimmon fruit stored at 0.5 °C and 95% relative humidity over a 3-month period. The treatments caused a reduction in weight loss; in the middle of the cold storage, the soluble solids content was higher with the AVG treatment than with the 1-MCP treatment. Both treatments reduced the incidence and severity of peel blackening and fruit softening, whereas fruit decay was only affected by AVG. The content of total phenolic compounds was lower with AVG than with 1-MCP or in the untreated control during the same period. At the end of cold storage, the AVG treatment resulted in a greater total flavonoid and total antioxidant activity and a higher reduced glutathione level than with 1-MCP or in the untreated control. AVG likely evoked the least physiological and biochemical responses in persimmons, according to the results of a normalized heatmap matrix system. Correlation coefficient networking results showed that 1-MCP and AVG caused differential correlation responses between the fruit quality variables and targeted metabolites during cold storage. Overall, the results indicated that treatment of cold-stored persimmons with 1-MCP and AVG likely reduced the incidence of physiological disorders, although antioxidant metabolites were relatively less affected by both regulators.

Sensory Evaluation of Dry Persimmons of the Tipo (Diospyros kaki L. f.) Variety

The study carried out at the Faculty of Agriculture and Life Sciences of the University of Maribor aimed to evaluate the attractiveness of dried persimmon fruit depending on the method of preparing the fruit for drying. The methods varied in terms of the ripeness of the fruit, the thickness and shape of the slices, the presence of seeds (in fertilised fruits) and the presence of peel. Sensory evaluation was performed by students and staff, using a hedonic scale, where evaluators evaluated the appearance (shape and colour) and taste (sugar-acid ratio, texture, tartness, presence of fruit peel, and general impression) of the fruit. Dried unfertilized persimmons, with the presence of peel, cut into 3 mm thick slices, were given the highest score for overall attractiveness. In terms of taste, unfertilised, peeled persimmon slices scored the highest. Considering all parameters, the results showed that unfertilised persimmons were more suitable for drying and that the presence of the peel was not a disturbing factor for consumers.

Chlorophyll fluorescence imaging as a tool to evaluate calyx senescence during the ripening of persimmon fruit treated with gibberellic acid

The effect of gibberellic acid (GA) on retarding loss of persimmon firmness and fruit coloration has been previously reported. Nevertheless, information about the effect of this treatment on calyx senescence is lacking. In this study, chlorophyll fluorescence imaging (CFI) was used to evaluate calyx senescence in persimmon fruit treated with GA. At three harvest times, physico-chemical parameters measured on persimmon fruit and CFI parameters (Fo, Fm and Fv/Fm) on calyx sepals were evaluated on the fruit treated once or twice with GA, and also on untreated fruit (CTL). A decline in the chlorophyll fluorescence parameters correlated with calyx senescence and progressed during fruit ripening. Spatial images heterogeneity in the Fm/Fv measurements illustrates senescence and necrosis dynamics, which began in the apical area of sepals and progressed to the basal area. Besides retarding fruit ripening, GA treatments delayed the calyx senescence process, and hence improved external fruit quality maintenance. The CFI parameters measured on the calyx correlated with both external color evolution and firmness loss during fruit ripening. Consequently, these chlorophyll fluorescence parameters could act as a potential non-intrusive tool to determine persimmon harvesting time.

Pesticide Inhalation Exposure of Applicators and Bystanders Using Conventional and Innovative Cropping Systems in the Valencian Region, Spain

This paper provides scientific results from a European LIFE project carried out in the Valencian region of Spain during the 2017 to 2018 time frame. In 2018, more than 60,000 tons of pesticides were commercialized in Spain, with approximately 15% destined for Valencian crops. In order to improve the air quality in the agricultural areas of this region, an innovative cropping system based on irrigation was developed and compared to conventional treatments based on hand-spray and turbo application. After applying conventional treatments to five types of crops (citrus, persimmon, nectarine, watermelon, and other stone fruits), a total of 13 active substances were detected in the air. The same active substances were applied to crops using the novel irrigation system, and no pesticide was detected in the air. Moreover, applicator and bystander populations in the region were assessed for their risk of inhalation exposure to pesticides, and no risk was found when either of the techniques, the innovative and the conventional agricultural one, were applied.

Microbial community composition during artificial frosting of dried persimmon fruits

Spoilage caused by microbial contamination in dried persimmon fruits processed by traditional frosting often leads to considerable economic losses. Artificial frosting has recently been considered as an alternative method to reduce the risk of spoilage, however, in this study, expansion of packaging and mildew formation were still observed. The dynamics of bacteria and fungi structure in different processes were determined. Results showed that Zygosaccharomyces and Aspergillus were the predominant genera in spoiled fruits in the packaging phase. During the artificial frosting, Wickerhamomyces (15.91%) and Candida (6.82%) were the predominant gas-generated yeasts and Aspergillus (20.45%) and Penicillium (15.91%) were consistently the dominant genera causing mildew. To reduce the contamination problem, pulsed light (24 J/cm2) were adopted on Day 0 of frosting and no fungi were detected on Day 3 post-frosting, which indicated to decrease significantly the amounts of Wickerhamomyces, Candida, Aspergillus, and Penicillium. Additionally, the results of the high-throughput sequencing suggested that the proportions of these predominant microorganisms were also decreased significantly during frosting. These results showed that pulsed light treatment in the artificial technique could almost overcome the problems of packaging expansion and mildew formation.